EDTA-functionalized magnetic nanoparticles: A suitable platform for the analysis of low abundance urinary proteins

TALANTA

卷:170

Article

Bastos, Paulo¹  Trindade, Fabio^1,2  Ferreira, Rita³  Leite-Moreira, Adelino²  Falcao-Pires, Ines²  Manadas, Bruno⁴  Daniel-da-Silva, Ana L.⁵  Vitorino, Rui^1,2 

[1] Univ Aveiro, IBiMED Inst Biomed, Dept Med Sci, Aveiro, Portugal

[2] Univ Porto, Dept Cirurgia & Fisiol, Unidade Invest Cardiovasc, Fac Med, Oporto, Portugal

[3] Univ Aveiro, Dept Chem, Mass Spectrometry Ctr, QOPNA, Aveiro, Portugal

[4] Univ Coimbra, CNC Ctr Neurosci & Cell Biol, Coimbra, Portugal

[5] Univ Aveiro, CICECO Aveiro Inst Mat, Dept Chem, Aveiro, Portugal

关键词: Magnetic Nanoparticles;    Proteomics;    Urine;    Fractionation;    Mass spectrometry;   

DOI  :  10.1016/j.talanta.2017.03.087

来源: Elsevier

【 摘 要 】

Urine is a highly attractive source of biological information and disease biomarkers, whose proteome characterization is ongoing. To that end, depletion/enrichment strategies for protein analysis can be of great convenience. We have thus developed a method based on the use of EDTA-functionalized magnetic nanoparticles (NPs@EDTA), to fractionate urine samples before liquid chromatography-mass spectrometry analysis and compared the identified proteins with those obtained from ultrafiltrated/unfractionated (UF) urine samples. NPs@EDTA allowed larger urine volumes to be processed, resulting in a greater number of protein identifications (similar to 2-fold) at a lower cost when compared to UF samples. Proteins of greater abundance (such as albumin and uromodulin) were, at least partially, depleted with NPs@EDTA while those of lower abundance were enriched. Bioinformatics analysis showed that approximately 27% of NPs@EDTA-enriched proteins were annotated as displaying enzymatic activity, most of these being hydrolytic enzymes (56%), particularly proteases/peptidases (48%). Also, post-translational modifications were prominently predicted across NPs@ EDTA-enriched proteins (90%), particularly glycosylation (52%), phosphorylation (47%) and acetylation (30%). NPs@EDTA allowed the identification of 109 proteins in urine for the first time, showing high potential as a platform for urine's fractionation prior to proteomic analysis.

【 授权许可】

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